Method of promoting cervical and vaginal secretions

ABSTRACT

The present invention provides a method of stimulating cervical and vaginal secretions in a mammal by treatment with P2Y 2  and/or P2Y 4  purinergic receptor agonists. Treatment of vaginal dryness associated with menopause, chemotherapy, and various disease states as well as the treatment of vulvar pain is discussed. Suitable agonists such as UTP, CTP, ATP, dinucleotides and analogs thereof are disclosed.

[0001] This application is a continuation-in-part of U.S. applicationSer. No. 09/122,516 filed Jul. 24, 1998, which claims priority to U.S.Provisional Application Ser. No. 60/054,147, filed Jul. 25, 1997. Bothapplications are incorporated herein by reference.

TECHNICAL FIELD

[0002] This invention relates to a method of regulating secretions inand around the cervix and vagina of a patient by administeringpurinergic receptor agonists such as certain uridine, adenine, orcytidine triphosphates as well as other nucleoside phosphate containingcompounds.

BACKGROUND OF THE INVENTION

[0003] Vaginal dryness is a very common problem which brings physicaland emotional distress to many women (Key, E., Nurs. Stand. 5:24-27(1991)). It most commonly manifests itself during sexual intercourse,which causes dyspareunia and can eventually lead to apareunia. Althoughit is traditionally considered to be a condition which affectspostmenopausal women, it can occur during the premenopausal andperimenopausal years. The use of oral contraceptives may also cause areduction in vaginal moisture in some women (Reginald, W., et al., Br.J. Obstet. Gynaecol. 96:1148-1152 (1989)). Postpartum vaginal dryness,independent of or as a result of lactation, can be a significantcomplaint (Wisniewski, P., et al., Am. J. Obstet. Gynecol. 165:1249-1254(1991)). Women undergoing chemotherapy or radiotherapy for malignantdiseases such as leukemia often experience vaginal dryness as a, resultof treatment (Cust, M., et al., Br. Med. J. 299:1494-1497 (1989)). Manydisease states, such as systemic sclerosis and other systemic autoimmunedisorders (Bhadauria, S., et al., Am. J. Obstet. Gynecol. 172:580-587(1995)), Ehlers-Danlos syndrome (Sorokin, Y., et al., J. Reprod. Med.39:281-284 (1994)), diabetes mellitus (Sreebny, L., et al., DiabetesCare 15:900-904 (1992)), and Sjögren's syndrome (Mdarchesoni, D., etal., Eur. J. Obstet. Gynecol. Reprod. Biol. 63:49-53 (1995)) havedecreased vaginal hydration and lubrication problems as significantdisease-asscociated symptoms.

[0004] Vulvar pain is defined as the excessive sensitivity of the nervessupplying the mucus membrane of the vulva. This persistent burning andsensitivity in vulvar skin is not caused by identifiable infection. Itcannot be cured by surgery. The diseases covered under “vulvar pain” arealso referred to as vulvodynia/vulvar vestibulitis, vulvitis, burningvulvar syndrome and is often associated with fibromylagia, irritablebowel syndrome, Sjögren's syndrome, chronic inflammation, and Paget'sdisease as well as in the absence of any identifiable disease orinfection. R. Paul St. Afrmad, M.D., an endocrinologist at UCLA, hassuccessfully treated fibromylagia with uricosuric (gout) drugs,especially guaifenesin, a drug used to liquefy mucus (Yount, J. J. etal., Women's Health Digest 3(2) 1997). Dr. Armad has found that suchgout drugs provide an effective treatment for fibromylagia, even thoughgout and fibromylagia have no connection. Dr. Armad has found that24-hour urine samples taken from patients before and after treatmentexhibited a significant increase in the excretion of phosphate and amoderate increase of oxalate and calcium after guaifenesin was started.His hypothesis is that an excess of intracellular phosphate, andpossibly oxalate, builds up in the cells of fibromylagia sufferers anddepresses formation of energy (ATP) in the mitochondria of the cells. Itshould be noted that the role of ATP in Dr. Armad's theory is as anenergy source and not an agonist of the P2Y₂ receptor.

[0005] Current therapies for increasing vaginal moisture are:lubricating agents such as lubricating creams or jellies, topicalestrogen creams, and HRT (hormone replacement therapy). Lubricatingjellies provide short-lived and temporary relief, as these are aqueouspreparations containing no pharmacologically active agent. Topicalestrogen creams, if used on a regular basis, may be absorbed into thesystemic circulation. This can cause endometrial stimulation and canlead to endometrial hyperplasia and carcinoma (Whitehead, M., et al., N.Eng. J. Med. 305:1599-1605 (1981)). HRT is effective at relievingsymptoms of vaginal atrophy and hence vaginal dryness but has severalcontraindications and unwanted risks and side effects. A history of gallbladder disease (N. Eng. J. Med., 290:15-19 (1974)) or a personal orfamily history of reproductive or breast cancer (Harlap, S., Am. J.Obstet. Gynecol. 166:1986-1992 (1992)) are contraindications forestrogen therapy. Other contraindications are: history of stroke,cardiovascular disease, deep-vein thrombosis, superficialtlhrombophlebitis, liver disease, heavy smoking, high blood pressure,diabetes, uterine bleeding or large fibroids, hyperlipidemia, and grossobesity (Lichtman, R., J. Nurse Midwifery 36:30-48 (1991)). One majordisadvantage of HRT is the resumption of monthly withdrawal bleeds,which many postmenopausal women will not accept. Some women, even whileon HRT, still experience a degree of vaginal dryness (Key, E., Nurs.Stand. 5:24-27 (1991)).

[0006] It has been shown that uridine 5′-triphosphate (UTP) anddinucleotides such as diuridine tetraphosphate are potent agonists ofP2Y₂ purinergic receptors found on the surface of human airwayepithelium. UTP has been shown to increase both the rate and totalamount of mucin secreted by goblet cells in vitro (Lethem, M., et al.,Am. J. Respir. Cell Mol. Biol. 9:315-322 (1993)). UTP has also beenshown to increase chloride secretion, and hence, water secretion fromairway epithelial cells in vitro (Mason, S., et al., Br. J. Pharmacol.103:1649-1656 (1991)).

[0007] Thus, as a result of the ineffectiveness and risks of currenttherapies, medical researchers have sought to develop alternatives forthe treatment of vaginal dryness. Because of the demonstrated ability ofUTP and dinucleotides, such as diuridine tetraphosphate, to increasehydration of airway epithelial secretions and stimulate release ofmucins, applicants were motivated to investigate whether UTP and otherP2Y₂ and/or P2Y₄ purinergic receptor agonists could stimulate hydrationand mucin production in the vaginal and cervical epithelia.

SUMMARY OF THE INVENTION

[0008] A method of stimulating cervical and vaginal secretions in asubject in need of such treatment is disclosed. The method of thepresent invention may be used to increase cervical and vaginalsecretions for any reason, including, but not limited to, treatment ofvaginal dryness and/or treatment of vulvar pain. Vaginal dryness isassociated with but not limited to menopause, childbirth, breastfeeding,chemotherapy or radiotherapy, diabetes mellitus, Sjögren's syndrome,Ehlers-Danlos syndrome, systemic sclerosis and other systemic autoimmunediseases, hysterectomy, urogenital surgery, psychosomatic disorders,anxiety, psychosexual problems, and pharmacological drug-related sideeffects. The method of the present invention comprises administering aP2Y₂ and/or P2Y₄ purinergic receptor agonist: uridine 5′-triphosphate,P¹,P⁴-di(uridine-5′)tetraphosphate, cytidine 5′-triphosphate oradenosine 5′-triphosplhate or analogs thereof, in an amount effective tostimulate vaginal and cervical secretions.

[0009] Another aspect of the present invention is the use of uridine5′-triphosphate, P¹,P⁴-di(uridine-5′)tetraphosphate, cytidine5′-triphosphate or adenosine 5′-triphosphate or analogs thereof, for themanufacture of a medicament for carrying out a therapeutic method oftreatment as given above.

[0010] The present invention also discloses pharmaceutical compositionscomprising uridine 5′-triphosphate, P¹,P⁴-di(uridine-5′)tetraphosphate,cytidine 5′-triphosphate or adenosine 5′-triphosphate or analogsthereof, with a pharmaceutical carrier therefor.

DETAILED DESCRIPTION OF THE INVENTION

[0011] Applicants have discovered that uridine 5′-triphosphate (UTP) andrelated compounds are potent agonists for purinergic receptors found incervical and vaginal epithelia preparations. The methods of the presentinvention are an improvement upon the current most commonly usedtreatments of vaginal dryness as UTP stimulates a patient's ownproduction and secretion of mucins as well as increasing the levels ofmucosal hydration, which serve to maintain the natural protective andlubricant characteristics of vaginal and cervical mucosa. The methods ofthe present invention may also be used exclusive of, or as an adjunctto, hormone replacement therapy (HRT) or estrogen replacement therapy(ERT).

[0012] The present invention provides a method of stimulating cervicaland vaginal secretions in a mammal, including a human, in need thereofby administering an amount of a compound of Formulas I, II, III, or IVor a pharmaceutically acceptable ester or salt thereof effective toincrease said secretions.

[0013] UTP and its analogs are depicted in general Formula I:

[0014] wherein:

[0015] X₁, X₂ and X₃ are each independently either O⁻ or S⁻; preferably,X₂ and X₃ are O⁻;

[0016] R₁ is O, imido, methylene or dihalomethylene (e.g.,dichloromethylene or difluoromethylene); preferably, R₁ is oxygen ordifluoromethylene;

[0017] R₂ is H or Br; preferably, R₂ is H; particularly preferredcompounds of Formula I are uridine 5′-triphosphate (UTP) and uridine5′-O-(3-thiotriphosphate) (UTPγS).

[0018] A dinucleotide is depicted by the general Formula II:

Formula II

[0019]

[0020] wherein:

[0021] X is oxygen, methylene, difluoromethylene, imido;

[0022] n=0, 1, or 2;

[0023] m=0, 1, or 2;

[0024] n+m=0, 1, 2, 3, or 4; and

[0025] B and B′ are each independently a purine residue or a pyrimidineresidue linked through the 9- or 1-position, respectively;

[0026] Z=OH or N₃;

[0027] Z′=OH or N₃;

[0028] Y=H or OH;

[0029] Y′=H or OH;

[0030] provided that when Z is N₃, Y is H or when Z′ is N₃, Y′ is H.

[0031] The furanose sugar is preferably in the β-configuration.

[0032] The faranose sugar is most preferably in the β-D-configuration.

[0033] Preferred compounds of Formula II are the compounds of FormulaIIa:

[0034] wherein:

[0035] X=O;

[0036] n+m=1 or 2;

[0037] Z, Z′, Y, and Y′=OH;

[0038] B and B′ are defined in Formulas IIc and IId;

[0039] X=O;

[0040] n+m=3 or 4;

[0041] Z, Z′, Y, and Y′=OH;

[0042] B=uracil;

[0043] B′ is defined in Formulas IIc and IId; or

[0044] X=O;

[0045] n+m=1 or 2;

[0046] Z, Y, and Y′=OH;

[0047] Z′=H;

[0048] B=uracil;

[0049] B′ is defined in Formulas IIc and IId; or

[0050] X=O;

[0051] n+m=0, 1, or 2;

[0052] Z and Y=OH;

[0053] Z′=N₃;

[0054] Y′=H;

[0055] B=uracil;

[0056] B′=thymine; or

[0057] X=O;

[0058] n+m=0, 1, or 2;

[0059] Z and Z′=N₃;

[0060] Y and Y′=H;

[0061] B and B′=thymine; or

[0062] X=CH₂, CF₂, or NH;

[0063] n and m=1;

[0064] Z, Z′, Y, and Y′=OH;

[0065] B and B′ are defined in Formulas IIc and lId.

[0066] Another preferred group of the compounds of Formula II are thecompounds of Formula IIb or the pharmaceutically acceptable saltsthereof:

[0067] wherein:

[0068] X is oxygen, methylene, difluoromethylene, or imido;

[0069] n=0 or 1;

[0070] m=0 or 1;

[0071] n+m=0, 1, or 2; and

[0072] B and B′ are each independently a purine residue, as in FormulaIIc, or a pyrimidine residue, as in Formula IId, linked through the 9-or 1-position, respectively. In the instance where B and B′ are uracil,attached at N-1 position to the ribosyl moiety, then the total of m+nmay equal 3 or 4 when X is oxygen. The ribosyl moieties are in the D-configuration, as shown, but may be L-, or D- and L-. The D-configuration is preferred.

[0073] The substituted derivatives of adenine include adenine 1-oxide;1,N6-(4- or 5-substituted etheno) adenine; 6-substituted adenine; or8-substituted aminoadenine, where R′ of the 6- or 8-HNR′ groups arechosen from among: arylalkyl (C₁₋₆) groups with the aryl moietyoptionally functionalized as described below; alkyl; and alkyl groupswith functional groups therein, such as:([6-aminohexyl]carbamoylmethyl)-, and ω-acylated-amino(hydroxy, thioland carboxy) derivatives where the acyl group is chosen from among, butnot limited to, acetyl, trifluroroacetyl, benzoyl, substituted-benzoyl,etc., or the carboxylic moiety is present as its ester or amidederivative, for example, the ethyl or methyl ester or its methyl, ethylor benzamido derivative. The co-amino(hydroxy, thiol) moiety may bealkylated with a C₁₋₄ alkyl group.

[0074] Likewise, B or B′ or both in Formula IIb may be a pyrimidine withthe general formula of Formula IId, linked through the 1-position:

[0075] wherein:

[0076] R₄ is hydroxy, mercapto, amino, cyano, aralkoxy, C₁₋₆ alkoxy,C₁₋₆ alkylamino, and dialkylamino, the alkyl groups optionally linked toform a heterocycle:

[0077] R₅ is hydrogen, acyl, C₁₋₆ alkyl, aroyl, C₁₋₅ alkanoyl, benzoyl,or sulphonate;

[0078] R₆ is hydroxy, mercapto, alkoxy, aralkoxy, C₁₋₆-alkylthio, C₁₋₅disubstituted amino, triazolyl, alkylamino, or dialkylamino, where thealkyl groups are optionally linked to form a heterocycle or linked toN-3 to form an optionally substituted ring;

[0079] R₇ is hydrogen, hydroxy, cyano, nitro, alkenyl, with the alkenylmoiety optionally linked through oxygen to form a ring optionallysubstituted on the carbon adjacent to the oxygen with alkyl or arylgroups, substituted alkynyl or hydrogen where R₈ is amino or substitutedamino and halogen, alkyl, substituted alkyl, perhalomethiyl (e.g., CF₃),C₂₋₆ alkyl, C₂₋₃ alkenyl, or substituted ethenyl (e.g., allylamino,bromvinyl and ethyl propenoate, or propenoic acid), C₂₋₃ alkynyl orsubstituted alkynyl when R₆ is other than amino or substituted amino andtogether R₅-R₆ may form a 5- or 6-membered saturated or unsaturated ringbonded through N or O at R₆, such a ring may contain substituents thatthemselves contain functionalities;

[0080] R₈ is hydrogen, alkoxy, arylalkoxy, alkylthio, arylalkylthio,carboxamidomethyl, carboxymethyl, methoxy, methylthio, phenoxy, orphenylthio.

[0081] In the general structure of Formula IId above, the dotted linesin the 2- to 6-positions are intended to indicate the presence of singleor double bonds in these positions; the relative positions of the doubleor single bonds being determined by whether the R₄, R₆, and R₇substituents are capable of keto-enol tautomerism.

[0082] In the general structures of Formula IIc and IId above, the acylgroups advantageously comprise alkanoyl or aroyl groups. The alkylgroups advantageously contain 1 to 8 carbon atoms, particularly 1 to 4carbon atoms optionally substituted by one or more appropriatesubstituents, as described below. The aryl groups including the arylmoieties of such groups as aryloxy are preferably phenyl groupsoptionally substituted by one or more appropriate substituents, asdescribed below. The above mentioned ailkenyl and alkynyl groupsadvantageously contain 2 to 8 carbon atoms, particularly 2 to 6 carbonatoms, e.g., ethenyl or ethynyl, optionally substituted by one or moreappropriate substituents as described below. Appropriate substituents onthe above-mentioned alkyl, alkenyl, alkynyl, and aryl groups areadvantageously selected from halogen, hydroxy, C₁₋₄ alkoxy, C₁₋₄ alkyl,C₆₋₁₂ arylalkoxy, carboxy, cyano, nitro, sulfonamido, sulfonate,phosphate, sulfonic, amino, and substituted amino wherein the amino issingly or doubly substituted by a C₁₋₄ alkyl, and when doublysubstituted, the alkyl groups optionally being linked to form aheterocycle.

[0083] For purposes of further clarifying the foregoing descriptions ofFormulae IIc and IId, the descriptions can be simplified to thefollowing:

[0084] R₂ is O or is absent; or

[0085] R₁ and R₂ taken together may form optionally substituted5-membered fused imidazole ring; or

[0086] R₁ of the 6-HNR₁ group or R₃ of the 8-HNR₃ group is chosen fromthe group consisting of:

[0087] (a) arylalkyl (C₁₋₆) groups with the aryl moiety optionallysubstituted,

[0088] (b) alkyl,

[0089] (c) ([6-aminohexyl]carbamoylmethyl),

[0090] (d) ω-amino alkyl (C₂₋₁₀),

[0091] (e) ω-hydroxy alkyl (C₂₋₁₀),

[0092] (f) ω-thiol alkyl (C₂₁₀),

[0093] (g) ω-carboxy alkyl (C₂₋₁₀),

[0094] (h) the ω-acylated derivatives of (b), (c) or (d) wherein theacyl group is either acetyl, trifluroacetyl, benzoyl, orsubstituted-benzoyl alkyl(C₂₋₁₀), and

[0095] (i) ω-carboxy alkyl (C₂₋₁₀) as in (e) above wherein thecarboxylic moiety is an ester or an amide;

Formula IId

[0096]

[0097] wherein:

[0098] R₄ is hydroxy, mercapto, amino, cyano, aralkoxy, C₁₋₆ alkylthio,C₁₋₆ alkoxy, C₁₋₆ alkylamino or dialkylamino, wherein the alkyl groupsof said dialkylamino are optionally linked to form a heterocycle;

[0099] R₅ is hydrogen, acyl, C₁₋₆ alkyl, aroyl, C₁₋₅ alkanoyl, benzoyl,or sulphonate;

[0100] R₆ is hydroxy, mercapto, alkoxy, aralkoxy, C₁₋₆-alkylthio, C₁₋₅disubstituted amino, triazolyl, alkylamino or dialkylamino, wherein thealkyl groups of said dialkylamino are optionally linked to form aheterocycle or linked to N³ to form an optionally substituted ring;

[0101] R₅-R₆ together forms a 5 or 6-membered saturated or unsaturatedring bonded through N or O at R₆, wherein said ring is optionallysubstituted;

[0102] R₇ is selected from the group consisting of:

[0103] (a) hydrogen,

[0104] (b) hydroxy,

[0105] (c) cyano,

[0106] (d) nitro,

[0107] (e) alkenyl, wherein the alkenyl moiety is optionally linkedthrough oxygen to form a ring optionally substituted with alkyl or arylgroups on the carbon adjacent to the oxygen,

[0108] (f) substituted alkynyl

[0109] (g) halogen,

[0110] (h) alkyl,

[0111] (i) substituted alkyl,

[0112] (j) perhalomethyl,

[0113] (k) C₂₋₆ alkyl,

[0114] (l) C₂₋₃ alkenyl,

[0115] (m) substituted ethenyl,

[0116] (n) C₂₋₃ alkynyl and

[0117] (o) substituted alkynyl when R₆ is other than amino orsubstituted amino;

[0118] R₈ is selected from the group consisting of:

[0119] (a) hydrogen,

[0120] (b) alkoxy,

[0121] (c) arylalkoxy,

[0122] (d) alkylthio,

[0123] (e) arylalkylthio,

[0124] (f) carboxamidomethyl,

[0125] (g) carboxymethyl,

[0126] (h) methoxy,

[0127] (i) methylthio,

[0128] (j) phenoxy and

[0129] (k) phenylthio.

[0130] CTP and its analogs are depicted by general Formula III:

Formula III

[0131]

[0132] wherein:

[0133] R₁, X₁, X₂ and X₃ are defined as in Formula I;

[0134] R₅ and R₆ are H while R₇ is nothing and there is a double bondbetween N-3 and C-4 (cytosine), or

[0135] R₅, R₆ and R₇ taken together are —CH═CH—, forming a ring from N-3to N-4 with a double bond between N-4 and C-4 (3,N⁴-ethenocytosine)optionally substituted at the 4- or 5-position of the etheno ring.

[0136] ATP and its analogs are depicted by general Formula IV:

[0137] wherein:

[0138] R₁, X₁, X₂, and X₃ are defined as in Formula I;

[0139] R₃ and R₄ are H while R₂ is nothing and there is a double bondbetween N-1 and C-6 (adenine), or

[0140] R₃ and R₄ are H while R₂ is O and there is a double bond betweenN-1 and C-6 (adenine 1-oxide), or

[0141] R₃, R₄, and R₂ taken together are —CH═CH—, forming a ring fromN-6 to N-1 with a double bond between N-6 and C-6 (1,N6-ethenoadenine).

[0142] For simplicity, Formulas I, II, III, and IV herein illustrate theactive compounds in the naturally occurring D-configuration, but thepresent invention also encompasses compounds in the L-configuration, andmixtures of compounds in the D- and L-configurations, unless otherwisespecified. The naturally occurring D-configuration is preferred.

[0143] The compounds of the invention may be present in the form oftheir pharmaceutically acceptable salts, such as, but not limited to, analkali metal salt such as sodium or potassium; an alkaline earth metalsalt such as manganese, magnesium, or calcium; or an ammonium ortetraalkyl ammonium salt, i.e., NX₄ ⁺ (wherein X is C₁₋₄).Pharrmaceutically acceptable salts are salts that retain the desiredbiological activity of the parent compound and do not impart undesiredtoxicological effects. The compounds of the invention may also bepresent in the form of prodrugs, typically comprising esters or amidemoieties on the heterocyclic and furanosyl hydroxyls of the compound.

[0144] Another aspect of the present invention is a method of treating amammal with vaginal dryness arising from, but not limited to, menopause,childbirth, breastfeeding, chemotherapy or radiotherapy, diabetesmellitus, Sjögren's syndrome, Ehlers-Danlos syndrome, systemic sclerosisand other systemic autoimmune diseases, hysterectomy, urogenitalsurgery, psychosomatic disorders, anxiety, psychosexual problems, andpharmacological drug-related side effects.

[0145] It is also contemplated that the method of the present inventioncan be used to increase vaginal moisture and lubrication in healthywomen for the purpose of facilitating sexual intercourse. It is furthercontemplated that the method of the present invention would beparticularly useful for a woman who wished to accommodate a sexualpartner who is undergoing treatment with Viagra® or other such drugsused for the treatment of erectile dysfunction.

[0146] The present invention further provides pharmaceuticalcompositions comprising a dosage form containing either P2Y₂ and/or P2Y₄purinergic receptor agonists selected from the group consisting ofgeneral Formula I, i.e., uridine 5′-triphosphate [UTP] and its analogs,general Formula II, i.e., P¹,P⁴-di(uridine-5′) tetraphosphate [U₂P₄] andits analogs, general Formula III, i.e., cytidine 5′-triphosphate [CTP]and its analogs, and general Formula IV, i.e., adenosine 5′-triphosphate[ATP] and its analogs.

[0147] The compounds disclosed herein may be administered to thecervical and/or vaginal mucosa of a patient by any suitable means, butare preferably administered by a solution, gel, suspension, cream, foam,pessary, or tablet containing the active compound. Alternatively, theactive compounds may by administered by continuous release from avaginal ring (Stumpf, P., Obstet. Gynecol. 75:9S (1990)) or anintrauterine device (Andersson, K., et al., Obstet. Gynecol. 79:963(1992)).

[0148] The topical solution, gel, jelly, ointment, cream, foam, pessary,or tablet contain the active compound in a physiologically compatiblevehicle, as those skilled in the art of gynecological topical deliverysystem development can select using conventional criteria.

[0149] Solutions formulated for administration to the vagina are usuallyreferred to as irrigations. These are sterile solutions, prepared in amanner typical of sterile injections that are intended for prepared as asingle use sterile solution.

[0150] Gels or jellies may be produced using a suitable gelling agentincluding, but not limited to, gelatin, tragacanth, or a cellulosederivative and may include glycerol as a humectant, emollient, andpreservative.

[0151] Ointments are semi-solid preparations that consist of the activeingredient incorporated into a fatty, waxy, or synthetic base.

[0152] Examples of suitable creams include, but are not limited to,water-in-oil and oil-in-water emulsions. Water-in-oil creams may beformulated by using a suitable emulsifying agent with propertiessimilar, but not limited, to those of the fatty alcohols such as cetylalcohol or cetostearyl alcohol and to emulsifying wax. Oil-in-watercreams may be formulated using an emulsifying agent such as cetomacrogolemulsifying wax. Suitable properties include the ability to modify theviscosity of the emulsion and both physical and chemical stability overa wide range of pH. The water soluble or miscible cream base may containa preservative system and may also be buffered to maintain an acceptablephysiological pH.

[0153] Foam preparations may be formulated to be delivered from apressurized aerosol canister, via a suitable applicator, using inertpropellants. Suitable excipients for the formulation of the foam baseinclude, but are not limited to, propylene glycol, emulsifying wax,cetyl alcohol, and glyceryl stearate. Potential preservatives includemethylparaben and propylparaben.

[0154] Pessaries are solid unit-dose forms suitably shaped for insertioninto the vagina and may either be composed of a base that melts at bodytemperature or which dissolves when in contact with mucous secretions.Examples of suitable bases include, but are not limited to, theobromaoil, synthetic fat bases (e.g. Witepsol), polyethylene glycols(macrogols), and glycerol suppository basis.

[0155] Vaginal tablets are composed of the active ingredient containedwithin a solid dosage form base which may include, but not be limitedto, excipients such as lactose, microcrystalline cellulose, corn starch,magnesium stearate, silicon dioxide, and hydroxypropyl methylcellulose.

[0156] In addition to the topical method of administration describedabove, there are various methods of administering the compounds of thepresent invention systemically. One such means would involve an aerosolsuspension of respirable particles comprised of the active compound,which the subject inhales. The active compound would be absorbed intothe bloodstream via the lungs and contact the cervical and/or vaginaltissues in a pharmaceutically effective amount. The respirable particlesmay be liquid or solid, with a particle size sufficiently small to passthrough the mouth and larynx upon inhalation; in general, particlesranging from about 1 to 10 microns, but more preferably 1-5 microns, insize are considered respirable.

[0157] Another means of systemically administering the active compoundsto the cervical and vaginal tissues of the subject would involveadministering a liquid/liquid suspension in the form of nasal drops of aliquid formulation, or a nasal spray of respirable particles which thesubject inhales. Liquid pharmaceutical compositions of the activecompound for producing a nasal spray or nasal drops may be prepared bycombining the active compound with a suitable vehicle, such as sterilepyrogen free water or sterile saline by techniques known to thoseskilled in the art.

[0158] Other means of systemic administration of the active compoundwould involve oral administration, in which pharmaceutical compositionscontaining compounds of Formulas I, II, III, or IV are in the form oftablets, lozenges, aqueous or oily suspensions, dispersible powders orgranules, emulsion, hard or soft capsules, or syrups or elixirs.Compositions intended for oral use may be prepared according to anymethod known to the art for the manufacture of pharmaceuticalcompositions and such compositions may contain one or more agentsselected from the group consisting of sweetening agents, flavoringagents, coloring agents, and preserving agents in order to providepharmaceutically elegant and palatable preparations. Tablets contain theactive ingredient in admixture with nontoxic pharmaceutically acceptableexcipients which are suitable for the manufacture of tablets. Theseexcipients may be, for example, inert diluents, such as calciumcarbonate, sodium carbonate, lactose, calcium phosphate, or sodiumphosphate; granulating and disintegrating agents, for example, cornstarch or alginic acid; binding agents, for example, starch, gelatin, oracacia; and lubricating agents, for example magnesium stearate, stearicacid, or talc. The tablets may be uncoated or they may be coated byknown techniques to delay disintegration and absorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonoistearate or glyceryl distearate may be employed. Formulations fororal use may also be presented as hard gelatin capsules wherein theactive ingredient is mixed with an inert solid diluent, for example,calcium carbonate, calcium phosphate, or kaolin, or as soft gelatincapsules wherein the active ingredient is mixed with water or an oilmedium, for example, peanut oil, liquid paraffin, or olive oil.

[0159] Additional means of systemic administration of the activecompound to the cervical and vaginal tissues of the subject wouldinvolve a suppository form of the active compound, such that atherapeutically effective amount of the compound reaches the cervicaland vaginal tissues via systemic absorption and circulation.

[0160] The quantity of the active compound included in thepharmaceutical composition is an amount sufficient to achieveconcentrations of the active compound on the cervical and/or vaginalmucosa of the subject of from about 10⁻⁷ to about 10⁻¹ Moles/liter, andmore preferably from about 10⁻⁶ to about 10⁻¹ Moles/liter.

[0161] Depending on the solubility of the particular formulation ofactive compound administered, the daily dose to promote cervical and/orvaginal mucin production and/or hydration may be divided among one orseveral unit dose administrations. The total daily dose for UTP (forexample) may range from 1 to 1000 milligrams, depending upon the age andstate of the subject, given at a regimen of up to four times per day oron an as needed basis to address acute exacerbations.

[0162] Some compounds of Formulas I, II, III, and IV can be made bymethods which are well known to those skilled in the art and inaccordance with known procedures (Zamecnik, P., et al., Proc. Natl Acad.Sci. USA 89:2370-2373 (1992); Ng, K., et al., Nucleic Acids Res.15:3572-3580 (1977); Jacobus, K. M., et al., U.S. Pat. No. 5,789,391 andPendergast, W., et al., International Patent Application WO98/34942));some are commercially available, for example, from Sigma ChemicalCompany, PO Box 14508, St. Louis, Mo. 63178. The synthetic methods ofU.S. Pat. No. 5,789,391 and International Patent Application WO98/34942are incorporated herein by reference.

EXAMPLES Example 1 In Vitro Short Circuit (I_(sc)) Measurements

[0163] The compound UTP is a potent agonist of P2Y₂ and/or P2Y₄purinergic receptors in cervical and vaginal tissue preparations byevaluation in vitro by administering UTP to the tissue culturesufficient to achieve concentrations of UTP on the mucosa of from about10⁻⁷ to about 10⁻¹ moles/liter. (Rojanasakul, Y., et al., Pharm. Res.9:1029-34 (1992); Bechgaard, E., et al., Int. J. Pharm. 106:237-242(1994); Gipson, I., et al., Biol. Reprod. 56:999-1011, (1997)).Specifically, ovariectomized female white albino New Zealand rabbits aresacrificed and vaginal tissue is removed. The tissue is mounted on asupporting ring and clamped in an Ussing chamber. I_(sc) is measured asflowing from the epithelial side to the serosal side of the tissue.Approximately half of this current corresponds to chloride movementthrough the membrane and hence, this is an accurate measure of thecorresponding fluid movement.

Example 2 In vivo Study in Rabbits

[0164] The compounds of the invention are evaluated in vivo byadministrating UTP, or any of the other P2Y₂ and/or P2Y₄ agonists of thepresent invention to an animal in an amount sufficient to achieveconcentrations of P2Y₂ and/or P2Y₄ agonist on the cervical and/orvaginal mucosa of the animal of from about 10⁻⁷ to about 10⁻¹moles/liter (Richardson, J., et al., Int. J. Pharm. 56:29-35 (1989)).Specifically, ovariectomized female white albino New Zealand rabbits aredosed with a P2Y₂ and/or P2Y₄ agonist such as any of the compounds ofthe present invention. A vaginal smear is then obtained with a cottonswab. The sample is appropriately prepared, an ELISA or a colorimetricdot blot method is run on the sample, and the relative amounts ofrepresentative cervical mucins are determined as compared tonon-ovariectomized controls. (Gipson, I. et al., Biol. Reprod.56:999-1011 (1997)).

Example 3 In vivo Study Using Ovarectomized Cynologous Monkeys

[0165] The compounds of the present invention are evaluated in vivo withan animal model of vaginal dryness as follows. Ovariectomized cynomolgusmonkeys are examined before treatment and graded subjectively using afabinal atrophy index. The animals were then dosed intravaginally with100 to 300 μl mist containing 10⁻² to 1 moles/liter P2Y₂ and/or P2Y₄agonist, such as any of the compounds of the present invention. After10, 20, 30, 60 and 90 minutes the animals are subjected to agynecological exam and graded by qualified medical professions with thevaginal atrophy index on a scale of 1 to 5, including a measurement offluid pH. (Hubbard, G. et al., Lab Animal Sci. 47, 36-39, (1997)).

What is claimed is:
 1. A method of stimulating cervical and vaginalsecretions in a mammal in need thereof by administering an effectivesecretion stimulating amount of a compound of Formulas I, II, III, orIV:

wherein: X₁, X₂ and X₃ are each independently either O⁻ or S⁻, R₁ is O,imido, methylene or dihalomethylene; R₂ is H or Br; preferably, R₂ is H;or

wherein: X is oxygen, methylene, difluoromethylene, imido; n=0, 1, or 2;m=0, 1, or 2; n+m=0, 1, 2, 3, or 4; and B and B′ are each independentlya purine residue or a pyrimidine residue linked through the 9- or1-position, respectively; Z=OH or N₃; Z′=OH or N₃; Y=H or OH; Y′=H orOH; provided that when Z is N₃, Y is H or when Z′ is N₃, Y′ is H; or

wherein: R₁, X₁, X₂ and X₃ are defined as in Formula I; R₅ and R₆ are Hwhile R₇ is nothing and there is a double bond between N-3 and C-4(cytosine), or R₅, R₆ and R₇ taken together are —CH═CH—, forming a ringfrom N-3 to N-4 with a double bond between N-4 and C-4(3,N⁴-ethenocytosine) optionally substituted at the 4- or 5-position ofthe etheno ring; or

wherein: R₁, X₁, X₂, and X₃ are defined as in Formula I; R₃ and R₄ are Hwhile R₂ is nothing and there is a double bond between N-1 and C-6(adenine), or R₃ and R₄ are H while R₂ is O and there is a double bondbetween N-1 and C-6 (adenine 1-oxide), or R₃, R₄, and R₂ taken togetherare —CH═CH—, forming a ring from N-6 to N-1 with a double bond betweenN-6 and C-6 (1,N6-ethenoadenine); or pharmaceutically acceptable estersor salts thereof.
 2. The method of claim 1 wherein the compounds ofFormula II are those of Formula IIa:

wherein: X=O; n+m=1 or 2; Z, Z′, Y, and Y′=OH; B and B′ are defined inFormulas IIc and IId:

R₂ is O or is absent; or R₁ and R₂ taken together may form optionallysubstituted 5-membered fused imidazole ring; or R₁ of the 6-HNR₁ groupor R₃ of the 8-HNR₃ group is chosen from the group consisting of: (a)arylalkyl (C₁₋₆) groups with the aryl moiety optionally substituted, (b)alkyl, (c) ([6-aminohexyl]carbamoylmethyl), (d) ω-amino alkyl (C₂₋₁₀),(e) ω-hydroxy alkyl (C₂₋₁₀), (f) ω-thiol alkyl (C₂₋₁₀), (g) ω-carboxyalkyl (C₂₋₁₀), (h) the ω-acylated derivatives of (b), (c) or (d) whereinthe acyl group is either acetyl, trifluroacetyl, benzoyl, orsubstituted-benzoyl alkyl(C₂₋₁₀), and (i) ω-carboxy alkyl (C₂₋₁₀) as in(e) above wherein the carboxylic moiety is an ester or an amide;

wherein: R₄ is hydroxy, mercapto, amino, cyano, aralkoxy, C₁₋₆alkylthio, C₁₋₆ alkoxy, C₁₋₆ alkylamino or dialkylarnino, wherein thealkyl groups of said dialkylamino are optionally linked to form aheterocycle; R₅ is hydrogen, acyl, C₁₋₆ alkyl, aroyl, C₁₋₅ alkanoyl,benzoyl, or sulphonate; R₆ is hydroxy, mercapto, alkoxy, aralkoxy,C₁₋₆-alkylthio, C₁₋₅ disubstituted amino, triazolyl, alkylarnino ordialkylamino, wherein the alkyl groups of said dialkylamino areoptionally linked to form a heterocycle or linked to N³ to form anoptionally substituted ring; R₅-R₆ together forms a 5 or 6-memberedsaturated or unsaturated ring bonded through N or O at R₆, wherein saidring is optionally substituted; R₇ is selected from the group consistingof: (a) hydrogen, (b) hydroxy, (c) cyano, (d) nitro, (e) alkenyl,wherein the alkenyl moiety is optionally linked through oxygen to form aring optionally substituted with alkyl or aryl groups on the carbonadjacent to the oxygen, (f) substituted alkynyl (g) halogen, (h) alkyl,(i) substituted alkyl, (j) perhalomethyl, (k) C₂₋₆ alkyl, (l) C₂₋₃alkenyl, (m) substituted ethenyl, (n) C₂₋₃ alkynyl and (o) substitutedalkynyl when R₆ is other than amino or substituted amino; R₈ is selectedfrom the group consisting of: (a) hydrogen, (b) alkoxy, (c) arylalkoxy,(d) alkylthio, (e) arylalkylthio, (f) carboxamidomethyl, (g)carboxymethyl, (h) methoxy, (i) methylthio, (j) phenoxy and (k)phenylthio wherein the substituted derivatives of adenine are adenine1-oxide; 1,N6-(4- or 5-substituted etheno) adenine; 6-substitutedadenine; or 8-substituted aminoadenine, where R′ of the 6- or 8-HNR′groups are chosen from among: arylalkyl (C₁₋₆) groups with the arylmoiety optionally functionalized; alkyl; and alkyl groups withfunctional groups therein, selected from the group consisting of([6-aminohexyl]carbamoylmethyl)-, and ω-acylated-amino(hydroxy, thioland carboxy) derivatives where the acyl group is acetyl,trifluroroacetyl, benzoyl or substituted-benzoyl and the carboxylicmoiety is present as the ethyl or methyl ester derivative or the methyl,ethyl or benzamido derivative; B or B′ or both in Formula IIb may be apyrimidine with the general formula of Formula IId, linked through the1-position:

wherein: R₄ is hydroxy, mercapto, amino, cyano, aralkoxy, C₁₋₆ alkoxy,C₁₋₆ alkylamino, and dialkylamino, the alkyl groups optionally linked toform a heterocycle; R₅ is hydrogen, acyl, C₁₋₆ alkyl, aroyl, C₁₋₅alkanoyl, benzoyl, or sulphonate; R₆ is hydroxy, mercapto, alkoxy,aralkoxy, C₁₋₆-alkylthio, C₁₋₅ disubstituted amino, triazolyl,alkylamino, or dialkylamino, where the alkyl groups are optionallylinked to form a heterocycle or linked to N-3 to form an optionallysubstituted ring; R₇ is hydrogen, hydroxy, cyano, nitro, alkenyl, withthe alkenyl moiety optionally linked through oxygen to form a ringoptionally substituted on the carbon adjacent to the oxygen with alkylor aryl groups, substituted alkynyl or hydrogen where R₈ is amino orsubstituted amino and halogen, alkyl, substituted alkyl, perhalomethyl,C₂₋₆ alkyl, C₂₋₃ alkenyl, or ethenyl (optionally substituted byallylamino, bromvinyl and ethyl propenoate, or propenoic acid), C₂₋₃alkynyl or substituted alkynyl when R₆ is other than amino orsubstituted amino and together R₅-R₆ may form a 5- or 6-memberedsaturated or unsaturated ring bonded through N or O at R₆, such a ringmay contain substituents that themselves contain functionalities; R₈ ishydrogen, alkoxy, arylalkoxy, alkylthio, arylalkylthio,carboxamidomethyl, carboxymethyl, methoxy, methylthio, phenoxy, orphenylthio; or X=O; n+m=3 or 4; Z, Z′, Y, and Y′=OH; B,=uracil; B′ isdefined in Formulas IIc and IId; or X=O; n+m=1 or 2; Z, Y, and Y′=OH;B=uracil; B′ is defined in Formulas IIc and IId; or X=O; n+m=0, 1, or 2;Z and Y=OH; Z′=N₃; Y′=H; B=uracil; B′=thymine; or X=O; n+m=0, 1, or 2; Zand Z′=N₃; Y and Y′=H; B and B′=thymine; or X=CH₂, CF₂, or NH; n andm=1; Z′, Z′, Y, and Y′=OH; B and B′ are defined in Formulas IIc and IId.3. The method of claim 1 wherein the compounds of Formula II are thoseof Formula IIb:

wherein: X is oxygen, methylene, difluoromethylene, or imido; n=0 or 1;m=or 1; n+m=0, 1, or 2; and B and B′ are each independently a purineresidue, as in Formula IIc as described in claim 2, or a pyrimidineresidue, as in Formula IId as described in claim 2, linked through the9- or 1-position, respectively; provided that when B and B′ are uracil,attached at N-1 position to the ribosyl moiety, then the total of m+nequals 3 or 4 when X is oxygen.
 4. The method of claim 1 wherein R₂ ofFormula I is H.
 5. The method of claim 1 wherein the furanose sugar ofFormula II is in the β-D-conifiguration.
 6. A method of treating amammal with vaginal dryness by administering an effective vaginaldryness treatment amount of a compound of Formulas I, II, III, or IV asdescribed in claims 1-5.
 7. A pharmaceutical composition comprising acompound of Formulas I, II, III, or IV as described in claims 1-5together with a pharmaceutically acceptable carrier therefor in the formof a liquid or gel suspension.
 8. The method of claim 6 wherein theamount of compound of Formulas I, II, III or IV administered to themammal is sufficient to achieve a concentration on the cervical and/orvaginal mucosa of from about 10⁻⁷ moles/liter to about 10⁻¹ moles/liter.9. The method of claim 6 wherein the amount of compound of Formulas I,II, III, or IV administered to the mammal is sufficient to achieve adaily dose of between 1 to 1000 milligrams.
 10. A method of treating amammal with vulvar pain by administrating an effective vulvar paintreatment amount of a compound of Formulas I, II, III, or IV asdescribed in claims 1-5.